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- W1004390544 abstract "Publisher Summary This chapter presents a method by which end-mature, intron-containing pre-tRNA is transcribed from a synthetic DNA template containing a bacteriophage T7 promoter. A Saccharomyces cerevisiae pre-tRNA Phe gene, containing a 19-base intron 4 and under the control of a bacteriophage T7 promoter, was constructed by self-assembly of eight synthetic oligodeoxynucleotides. The resulting DNA fragment was cloned into the Eco RI/ Bam HI site of the phage vector M13mpl0 RF. Double-stranded plasmid DNA containing the pre-tRNA gene is prepared in the usual way, except that CsCl ultracentrifugation is always done twice to ensure the cleanliness of the DNA template. The in vivo method precludes design and construction of site-specific mutations in the pre-tRNA gene, is laborious, and involves working with high levels of radioactivity. In the yeast nuclear extract (YNE) method, transcription and end processing take place simultaneously in the test tube. However, certain sequences in the mature domain of the pre-tRNA are known to affect these steps as well as the stability of the product." @default.
- W1004390544 created "2016-06-24" @default.
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- W1004390544 date "1989-01-01" @default.
- W1004390544 modified "2023-09-27" @default.
- W1004390544 title "[6] In vitro synthesis of end-mature, intron-containing transfer RNAs" @default.
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- W1004390544 doi "https://doi.org/10.1016/0076-6879(89)80092-8" @default.
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